LOCUS       Exported                8244 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Chloroplast transformation vector pAtTMT, complete sequence.
ACCESSION   JX235341
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8244)
  AUTHORS   Lu Y, Rijzaani H, Karcher D, Ruf S, Bock R.
  TITLE     Efficient engineering of the vitamin E metabolic pathway in 
            transgenic tobacco and tomato plastids with synthetic multigene 
            operons
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8244)
  AUTHORS   Lu Y, Rijzaani H, Karcher D, Ruf S, Bock R.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-JUN-2012) Organelle Biology, Biotechnology and 
            Molecular Ecophysiology, Max Planck Institute of Molecular Plant 
            Physiology, Am Muehlenberg 1, Potsdam-Golm, Brandenburg 14476, 
            Germany
REFERENCE   3  (bases 1 to 8244)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8244
                     /organism="Chloroplast transformation vector pAtTMT"
                     /mol_type="other DNA"
                     /db_xref="taxon:1229079"
     tRNA            complement(31..104)
                     /product="tRNA-Met"
     CDS             complement(254..556)
                     /codon_start=1
                     /product="ribosomal protein S14"
                     /label=ribosomal protein S14
                     /protein_id="AFQ99301.1"
                     /translation="MARKSLIQREKKRQKLEQKYHSIRRSSKKEISKVPSLSDKWEIYG
                     KLQSPPRNSAPTRLHRRCFLTGRPRANYRDFGLSGHILREMVHACLLPGATRSSW"
     gene            complement(679..1878)
                     /gene="psaB"
                     /label=psaB
                     /note="derived from Nicotiana tabacum"
     promoter        complement(1891..1909)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1919..1935)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      2077..2532
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2558..2662
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2663..3523
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             2861..3523
                     /codon_start=1
                     /gene="ampR"
                     /product="beta-lactamase"
                     /label=ampR
                     /protein_id="AFQ99302.1"
                     /translation="MSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHL
                     TDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEA
                     IPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFI
                     ADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     gene            2861..3523
                     /gene="ampR"
                     /label=ampR
     rep_origin      3618..4300
     rep_origin      3694..4282
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4570..4591
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4606..4636
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4644..4660
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     4668..4684
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4705..4723
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     gene            4737..4867
                     /gene="psbZ"
                     /label=psbZ
                     /note="derived from Nicotiana tabacum"
     tRNA            5143..5213
                     /product="tRNA-Gly"
     misc_recomb     5406..5439
                     /label=LoxP
                     /note="LoxP"
     protein_bind    complement(5406..5439)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      5480..5614
                     /regulatory_class="promoter"
                     /note="prrN promoter; derived from Nicotiana tabacum"
     CDS             5615..6406
                     /codon_start=1
                     /gene="aadA"
                     /product="streptomycin/spectinomycin adenylyltransferase"
                     /label=aadA
                     /protein_id="AFQ99303.1"
                     /translation="MAEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVLEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     gene            5615..6406
                     /gene="aadA"
                     /label=aadA
     regulatory      6417..6811
                     /regulatory_class="terminator"
                     /note="psbA terminator; derived from Nicotiana tabacum"
     misc_recomb     6815..6848
                     /label=LoxP
                     /note="LoxP"
     protein_bind    complement(6815..6848)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     primer_bind     6878..6894
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(6897..7117)
                     /regulatory_class="terminator"
                     /note="rbcL terminator; derived from Nicotiana tabacum"
     CDS             complement(7118..8026)
                     /codon_start=1
                     /gene="TMT"
                     /product="gamma-tocopherol methyltransferase"
                     /label=TMT
                     /note="derived from Arabidopsis thaliana"
                     /protein_id="AFQ99304.1"
                     /translation="MVAVAAAATSTEALRKGIAEFYNETSGLWEEIWGDHMHHGFYDPD
                     SSVQLSDSGHKEAQIRMIEESLRFAGVTDEEEEKKIKKVVDVGCGIGGSSRYLASKFGA
                     ECIGITLSPVQAKRANDLAAAQSLAHKASFQVADALDQPFEDGKFDLVWSMESGEHMPD
                     KAKFVKELVRVAAPGGRIIIVTWCHRNLSAGEEALQPWEQNILDKICKTFYLPAWCSTD
                     DYVNLLQSHSLQDIKCADWSENVAPFWPAVIRTALTWKGLVSLLRSGMKSIKGALTMPL
                     MIEGYKKGVIKFGIITCQKPL"
     gene            complement(7118..8026)
                     /gene="TMT"
                     /label=TMT
                     /note="AtTMT"
     regulatory      complement(8027..8178)
                     /regulatory_class="promoter"
                     /note="prrN promoter; derived from Nicotiana tabacum"
     RBS             8034..8056
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(8191..8209)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"